Fig. 1

Hypoxia-induced changes of IL-17A and ER stress markers in retinas and macrophages. a Western blotting analysis of GRP78, ATF-4 and IL-17A protein levels at P12, P15, P17, P21 in retinas of OIR mice compared with their age-matched controls. b Immunofluorescent staining of F4/80 (red, specific expressed by macrophage) and ATF-4 (green), GRP78 (green), IL-17A (green) on representative sections of retinas from P17 OIR mice and normal controls. Boxed areas are magnified in the images. Yellow represents co-localization of GRP78, ATF-4 or IL-17A and macrophages infltration. Scale bars, 100 μm. c, d Analysis of mRNA (c) and protein (d) levels of GRP78, ATF-4 and IL-17A in macrophages exposed to hypoxia for 0, 4, 6, 8, 12 or 24 h. e Immunofluorescent staining of F4/80 (red) and ATF-4 (green), GRP78 (green), IL-17A (green) in macrophages cultured under hypoxic conditions for 0 and 24 h. Yellow represents cellular localization of GRP78, ATF-4 or IL-17A. Scale bars, 50 μm. β-actin served as an endogenous reference for normalization. Data are shown as mean ± SEM, n = 6–8 per group for Real-time RT-PCR, n = 3 per group for western blotting. Each experiment repeated three times. ns, no significance. * P < 0.05 and **P < 0.01 compared with control groups. ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer